title: In Vitro Analysis of CsA-Induced Hepatotoxicity in HepG2 Cell Line: Oxidative Stress and α2 and β1 Integrin Subunits Expression.
abstract: BACKGROUND
  Cyclosporine A (CsA)-induced hepatotoxicity could be due to a reduction in α2β1 integrin expression that may either be from the direct effect of CsA itself or from reactive oxygen species (ROS) overproduction.
  
  
  OBJECTIVE
  In this study we aimed to identify the cellular mechanisms underlying CsA-induced hepatic injury by investigating the activation patterns of the antioxidant enzymes, using HepG2 as an in vitro model.
  
  
  METHODS
  HepG2 cells were cultured with different concentrations of CsA (0, 0.1, 1, 10 μg/ml) for 72 h. Effect of CsA on, 1) cellular integrity, 2) glutathione reductase (GR) and glutathione peroxidase (GPx) activity, 3) cellular levels of glutathione (GSH), 4) intracellular ROS, 5) ALT and AST activities, 6) urea production and 7) α2β1 integrin expression were assayed.
  
  
  RESULTS
  CsA treatment demonstrated a dose dependent increase in intracellular levels of ROS, GPx activity and decrease in GSH levels (P<0.05). GR activity was mildly attenuated in 1 and 10 µg/ml concentrations of CsA. Alanine aminotranferase (ALT) and aspartate aminotransferase (AST) levels increased in CsA treated cells, while urea synthesis was significantly decreased following treatment with higher concentrations of CsA (P<0.05). Significant down-regulation of β1integrin expression was observed in 1 and 10 µg/ml CsA treated cells while α2 integrin mRNA was significantly down-regulated in all CsA treated cells.
  
  
  CONCLUSIONS
  The observed reduction of α2β1 integrin expression following CsA treatment could be proposed as a possible pathway of CsA-induced hepatotoxicity. Further studies are required to elucidate whether this attenuated expression is due to the direct effect of CsA or caused by overproduction of ROS.
authors.affiliation: Biochemistry Department, Medical School, Shiraz University of Medicinal Sciences, Shiraz, IR Iran ; Recombinant Protein Laboratory, School of Advanced Medicinal Sciences and Technologies, Shiraz University of Medicinal Sciences, Shiraz, IR Iran.
authors.forename: Zohreh
authors.initials: Z
authors.lastname: Mostafavi-Pour
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Khademi
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Zal
authors.forename: Ahmad Reza
authors.initials: AR
authors.lastname: Sardarian
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Amini
keywords: Antioxidant; Cyclosporine; Enzymes; Integrin alpha2beta1